LOCUS       sequence-258312-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Engineered sgRNA-LEA3 (2 BoxB aptamers) generator circuit including 
            promoter Plux2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3007)
  AUTHORS   Liu Y, Wan X, Wang B
  TITLE     Engineered CRISPRa enables programmable eukaryote-like gene 
            activation in bacteria.
  JOURNAL   Nat Commun. 2019 Aug 26;10(1):3693. doi: 10.1038/s41467-019-11479-0.
  PUBMED    31451697
REFERENCE   2  (bases 1 to 3007)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-08-26"; volume: "10(1):3693. doi"; pages: " 
            10.1038/s41467-019-11479-0"
COMMENT     Sequence Label: pLY105
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..59
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    183..201
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    238..256
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     terminator      complement(365..392)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        400..434
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             446..457
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1396..1498
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1499..2158
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1565..1584)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2179..2273
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2387..2932
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2833..2852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg
       61 aagcggaata tatccctagg cctagtagga ccaaaacgaa aaaaggcccc cctttcggga
      121 ggcctctttt ctggaatttg gtaccgagaa gcttcaaaaa aagcaccgac tcggtgccac
      181 ttgggccctt cttcagggcc caagttgata acggactagc cttatttgaa cttgctaggg
      241 cccttcttca gggccctagc tctcaaaccg cgatcactgg aaataactct ctagttttat
      301 tcgaaagtaa caaaccattt tcttgcgtaa acctgtacga tcctacaggt ctctagtata
      361 taaacgcaga aaggcccacc cgaaggtgag ccagtgtgat tgacagctag ctcagtccta
      421 gggattgtgc tagctactag agattaaaga ggagaaatac tagatgaaaa acataaatgc
      481 cgacgacaca tacagaataa ttaataaaat taaagcttgt agaagcaata atgatattaa
      541 tcaatgctta tctgatatga ctaaaatggt acattgtgaa tattatttac tcgcgatcat
      601 ttatcctcat tctatggtta aatctgatat ttcaatccta gataattacc ctaaaaaatg
      661 gaggcaatat tatgatgacg ctaatttaat aaaatatgat cctatagtag attattctaa
      721 ctccaatcat tcaccaatta attggaatat atttgaaaac aatgctgtaa ataaaaaatc
      781 tccaaatgta attaaagaag cgaaaacatc aggtcttatc actgggttta gtttccctat
      841 tcatacggct aacaatggct tcggaatgct tagttttgca cattcagaaa aagacaacta
      901 tatagatagt ttatttttac atgcgtgtat gaacatacca ttaattgttc cttctctagt
      961 tgataattat cgaaaaataa atatagcaaa taataaatca aacaacgatt taaccaaaag
     1021 agaaaaagaa tgtttagcgt gggcatgcga aggaaaaagc tcttgggata tttcaaaaat
     1081 attaggttgc agtgagcgta ctgtcacttt ccatttaacc aatgcgcaaa tgaaactcaa
     1141 tacaacaaac cgctgccaaa gtatttctaa agcaatttta acaggagcaa ttgattgccc
     1201 atactttaaa aattaataac actgatagtg ctagtgtaga tcactactag agctcggtac
     1261 caaagacgaa caataagacg ctgaaaagcg tcttttttcg ttttggtccc tctagtatta
     1321 attttgtctt catgcatgaa gacaactagg ctctagaagc ggccgcgaat tagatatctg
     1381 gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag
     1441 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
     1501 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     1561 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     1621 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     1681 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
     1741 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1801 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     1861 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1921 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1981 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     2041 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     2101 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     2161 tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     2221 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     2281 ctagtaacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
     2341 gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg
     2401 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
     2461 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
     2521 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
     2581 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
     2641 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
     2701 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
     2761 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
     2821 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
     2881 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
     2941 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg
     3001 ccccgtt
//
